|Prenylation||Sumoylation||Tyrosine sulfation||Tyrosine nitration|
1.If the antigen is provided by the customer: modified peptide (>15 aa >90% purity, 15 mg) and unmodified control (>15 aa >90% purity, 11 mg).
2.The success rate of the modified antibody preparation is lower than the normal one. To ensure the probability of obtaining a useful antibody after immunization, we recommend to immune 3-4 rabbits according to our years of accumulated experience.
Modified peptide synthesis usually with a length of about 20 amino acids.Approximately 10mg are used in modified epitope affinity purification, 10mg used in antibody production, and 10mg used in QC.
Approximately 10mg non-modified peptide (about 20 amino acids) used in affinity purification.
To obtain a more specific modified antibody, sometimes we use shortened modified peptides to purify.
Proper design of a suitable antigen is the most critical step of getting an useful antibody.
We choose the best region to synthesis peptide coupling KLH as antigen to immune animals through professional analysis.
Each synthesized peptide goes through HLPC and MS tests to ultimately obtain a useful antibody.
ABclonal offers DNA methylation, RNA methylation, pan-acetylation, non-histone modification and other custom specific modified antibodies that are developed to meet your research needs.
Use Nipponbare callus (same quantity, about 3g) as sample material of ChIP experiment, 02g and 03g as target gene. Conduct IP experiment using Anti- H3K27me3 (ABclonal) and Anti- H3K27me3 (Company M).
The efficiency of abclonal anti- H3K27me3 immunoprecipitation is 10 times that of company M anti- H3K27me3.
Obtaining a useful site-specific antibody is not easy and most manufacturer can not achieve because of the antibody validation. ABclonal has built advanced testing platforms for Western blot, immunohistochemistry, and immunofluorescence-based assays, supplemented by immunoprecipitation, RIP, ChIP ,ChIP- seq, and shRNA knock- out validation. In additional to manufacture, we dedicate to be the best validation platform in the world. We take every measure to provide the most accurate and reliable data for each of our customers.
ABclonal's standard option for polyclonal antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular among researchers.
The protocol uses two New Zealand White rabbits that are Specific Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA).We guarantee that a single antigen will be injected into both animals.
|Control Serum Collection||Day 0||Pre-immune bleed (5mL per rabbit)|
|Primary Injection||Day 1||Immunize with 0.50mg of antigen in CFA, 10 SQ sites|
|1st Booster||Day 14||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|2nd Booster||Day 28||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 35||Bleed (~25mL per rabbit)|
|3rd Booster||Day 42||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 56, 58||Two bleeds (~50mL total per rabbit)|
|ELISA Test||Day 60|| ELISA titration (results available online);
Verify disposition of rabbits; decide to continue or terminate
|Purification||Optional||6-10 mg purified antibodies|